The reported relaxing effect of CO on various smooth muscle tissues could also be found in guinea pig ileal strips. The effect was pronounced after precontraction with 10-100 nM acetylcholine and rather small with KCl. Based on the photoreversibility of the CO-dependent relaxation, a photochemical action spectrum was established which showed a maximum at around 422 nm. This definitely rules out the participation of a cytochrome P450 dependent process as postulated for the CO induced relaxation of lamb ductus arteriosus. With regard to the potency of KCN and antimycin A to relax ileal smooth muscle, the involvement of respiratory chain inhibition was reinvestigated, but no indication for such a mechanism could be obtained. In analogy to the mechanism of CO-inhibition of platelet activation we found that CO about doubles cGMP levels in guinea pig ileal strips. This is similar to NO which also leads to effective relaxation. We propose that CO can be considered and experimentally used as a convenient activator of soluble G-cyclase in smooth muscle and platelets.